Difference between revisions of "Part:BBa K2213008"
Line 17: Line 17:
 
<partinfo>BBa_K2213008 parameters</partinfo>
 
<partinfo>BBa_K2213008 parameters</partinfo>
 
<!-- -->
 
<!-- -->
 +
Function:
 +
<br>
 +
This part is an improved version of https://parts.igem.org/Part:BBa_K562001, originally submitted by team Dundee 2011. The original part contained an illegal XbaI site which has been removed to make it biobrick compatible. This part has been expressed under different strength promoters. A high strength Anderson promoter here, low strength Anderson promoter (https://parts.igem.org/Part:BBa_K2213006), and a medium strength Anderson promoter (https://parts.igem.org/Part:BBa_K2213007).<br>
 +
<br>
 +
The PduD tag was combined with the high strength Anderson promoter (https://parts.igem.org/Part:BBa_J23104) and mCherry.<br>
 +
<br>
 +
https://static.igem.org/mediawiki/2017/e/e4/PromoterComparison800p.png<br>
 +
<br>
 +
A gradient of fluorescence is evident when compared to low and medium promoters.<br>
 +
<br>
 +
<br>
 +
<br>
 +
https://static.igem.org/mediawiki/2017/b/bd/TagExpression800p.jpeg
 +
<br>
 +
The expression levels compared to low and medium are as shown.<br>
 +
<br>
 +
<br>
 +
<br>
 +
https://static.igem.org/mediawiki/2017/1/12/HighPromoterComparison800p.png<br>
 +
<br>
 +
When expressed alone the distribution of the mCherry signal was homogeneous and fluorescence level was relatively high. In the presence of EutS the mCherry signal was less homogeneous but lacked obvious localisation, suggesting EutS isn’t enough to form proper BMCs. In the presence of EutSMN the mCherry signal clumped together, indicating localisation to the BMC.

Revision as of 20:43, 30 October 2017


HighPromoter_PduD(1-20)_mCherry

This part is an improved version of https://parts.igem.org/Part:BBa_K562001, submitted by Dundee 2011.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 8
    Illegal NheI site found at 31
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Function:
This part is an improved version of https://parts.igem.org/Part:BBa_K562001, originally submitted by team Dundee 2011. The original part contained an illegal XbaI site which has been removed to make it biobrick compatible. This part has been expressed under different strength promoters. A high strength Anderson promoter here, low strength Anderson promoter (https://parts.igem.org/Part:BBa_K2213006), and a medium strength Anderson promoter (https://parts.igem.org/Part:BBa_K2213007).

The PduD tag was combined with the high strength Anderson promoter (https://parts.igem.org/Part:BBa_J23104) and mCherry.

PromoterComparison800p.png

A gradient of fluorescence is evident when compared to low and medium promoters.



TagExpression800p.jpeg
The expression levels compared to low and medium are as shown.



HighPromoterComparison800p.png

When expressed alone the distribution of the mCherry signal was homogeneous and fluorescence level was relatively high. In the presence of EutS the mCherry signal was less homogeneous but lacked obvious localisation, suggesting EutS isn’t enough to form proper BMCs. In the presence of EutSMN the mCherry signal clumped together, indicating localisation to the BMC.